Dear all,
I,m trying to source a modified Trichrome reference for the technique I'll
outline below. I will most likely be inaccurate in timing or even sequence
of steps, as it is twenty years since I've used it and even then all we had
on our method cards was a hand written procedure without reference. It is
my intention to investigate this and other methods with emphasis on
sucesssful mordanting. If you recognise it could you please advise me of a
reference be it text or Journal. Because use is made of Gomori's one step
Trichrome solution, I suspect that this procedure only appeared after 1950.
If you have any personal experience, your comments would also be greatly
appreciated as I believe it was capricious in inexperienced hands, at least
mine were at the time.
Name of Method:- unknown.
Fixation:- Any.
Technique:-
1) Take sections to water.
2) Mix 1 part 10% HCl in Absolute alcohol with 2-3 parts 3% Potassium
Dichromate, and flood sections for 10-15 mins.
3) Rinse briefly with water. Mix equal parts of Weigert's A&B and flood
section for 6-10mins.
4)Briefly rinse with 80% alcohol then water.
5)Flood sections with Picro-Orange G for 3-5mins.(Formula now unknown)
6)Rinse in running water until only erythrocytes retain Orange G.
7) Apply Gomori's One Step Trichrome solution for 10 mins.
8) Drain and apply 0.5% Acetic acid for 1-2 mins.
9) Rinse in water, dehydrate etc.
Results:- Nuclei- blue black; collagen- blue or green depending upon
preference; muscle- red; erythrocytes- orange.
Regards Mike Rentsch (Downunder)
Traralgon. Vic. Aust.